Antisteroidal compounds and steroid withdrawal down-regulate cadherin-11 mRNA and protein expression levels in human endometrial stromal cells undergoing decidualisation in vitro

1999 ◽  
Vol 53 (4) ◽  
pp. 384-393 ◽  
Author(s):  
George T.C. Chen ◽  
Spiro Getsios ◽  
Colin D. MacCalman
Keyword(s):  
Protein Expression ◽  
Stromal Cells ◽  
Steroid Withdrawal ◽  
Endometrial Stromal Cells ◽  
Expression Levels ◽  
Mrna And Protein Expression

scholarly journals Regulation of A Disintegrin And Metalloproteinase with ThromboSpondin Repeats-1 Expression in Human Endometrial Stromal Cells by Gonadal Steroids Involves Progestins, Androgens, and Estrogens

2006 ◽  
Vol 91 (12) ◽  
pp. 4825-4835 ◽  
Author(s):  
Jiadi Wen ◽  
Hua Zhu ◽  
Shuko Murakami ◽  
Peter C. K. Leung ◽  
Colin D. MacCalman
Keyword(s):  
Protein Expression ◽  
Stromal Cells ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Endometrial Stromal Cells ◽  
Expression Levels ◽  
Mrna And Protein Expression ◽  
Regulatory Effects ◽  
A Disintegrin And Metalloproteinase

Abstract Context: Gonadal steroids are key regulators of the extracellular matrix remodeling events that occur in the human endometrium during each menstrual cycle. The spatiotemporal expression of A Disintegrin And Metalloproteinase with ThromboSpondin repeats (ADAMTS)-1 in human endometrial stroma in vivo suggests that this novel metalloproteinase may contribute to this tightly regulated developmental process. Objective: The objective of the study was to determine whether progesterone (P4), 17β-estradiol (E2), or the nonaromatizable androgen dihydrotestosterone (DHT), alone or in combination, is capable of regulating ADAMTS-1 mRNA and protein levels in human endometrial stromal cells in a concentration- and time-dependent manner. Design: A real-time quantitative PCR strategy and Western blotting were used to examine ADAMTS-1 mRNA and protein expression levels in primary cultures of human endometrial stromal cells. Results: P4 and DHT but not E2 increased the levels of the ADAMTS-1 mRNA transcript and protein species (110 kDa) present in endometrial stromal cells in vitro in a concentration- and time-dependent manner. A combination of P4 and DHT resulted in an additional increase in stromal ADAMTS-1 expression, whereas E2 attenuated the regulatory effects of P4 and DHT in a concentration-dependent manner. The antisteroidal compounds, mifepristone (RU486) and hydroxyflutamide, were also found to inhibit specifically the P4- and DHT-mediated increase in ADAMTS-1 mRNA and protein expression levels in these primary cell cultures in a concentration-dependent manner, respectively. Conclusions: These studies demonstrate that progestins, androgens, and estrogens, alone and in combination, have distinct regulatory effects on ADAMTS-1 mRNA and protein expression levels in human endometrial stromal cells in vitro.

scholarly journals MicroRNA-204-5p regulates apoptosis by targeting Bcl2 in rat ovarian granulosa cells exposed to cadmium†

2020 ◽  
Vol 103 (3) ◽  
pp. 608-619
Author(s):  
Ping Zhong ◽  
Jin Liu ◽  
Hong Li ◽  
Senbin Lin ◽  
Lingfeng Zeng ◽  
...  
Keyword(s):  
Protein Expression ◽  
Granulosa Cells ◽  
B Cell Lymphoma ◽  
Experimental Conditions ◽  
Expression Levels ◽  
Ovarian Granulosa Cells ◽  
Mrna And Protein Expression ◽  
Induced Apoptosis ◽  
Apoptosis Regulation

Abstract This study aimed to investigate whether cadmium (Cd) cytotoxicity in rat ovarian granulosa cells (OGCs) is mediated through apoptosis or autophagy and to determine the role of microRNAs (miRNAs) in Cd cytotoxicity. To test this hypothesis, rat OGCs were exposed to 0, 10, and 20 μM CdCl2 in vitro. As the Cd concentration increased, OGC apoptosis increased. In addition, Cd promoted apoptosis by decreasing the mRNA and protein expression levels of inhibition of B-cell lymphoma 2 (Bcl2). However, under our experimental conditions, no autophagic changes in rat OGCs were observed, and the mRNA and protein expression levels of the autophagic markers microtubule-associated protein 1 light chain 3 alpha (Map1lc3b) and Beclin1 (Becn1) were not changed. Microarray chip analysis, miRNA screening, and bioinformatics approaches were used to further explore the roles of apoptosis regulation-related miRNAs. In total, 19 miRNAs putatively related to Cd-induced apoptosis in rat OGCs were identified. Notably, miR-204-5p, which may target Bcl2, was identified. Then, rat OGCs were cultured in vitro and used to construct the miR-204-5p-knockdown cell line LV2-short hairpin RNA (shRNA). LV2-shRNA cells were exposed to 20 μM Cd for 12 h, and the mRNA and protein expression levels of Bcl2 were increased. Our findings suggest that Cd is cytotoxic to rat OGCs, and mitochondrial apoptosis rather than autophagy mediates Cd-induced damage to OGCs. Cd also affects apoptosis-related miRNAs, and the underlying apoptotic mechanism may involve the Bcl2 gene.

scholarly journals Tert-Butylhydroquinone Prevents Oxidative Stress-Mediated Apoptosis and Extracellular Matrix Degradation in Rat Chondrocytes

2021 ◽  
Vol 2021 ◽  
pp. 1-12
Author(s):  
Bo Yang ◽  
Haisheng Huang ◽  
Qisong He ◽  
Wei Lu ◽  
Lu Zheng ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Extracellular Matrix ◽  
Matrix Metalloproteinase ◽  
Protein Expression ◽  
Cell Viability ◽  
Cell Counting ◽  
Expression Levels ◽  
Extracellular Matrix Components ◽  
Mrna And Protein Expression

Oxidative stress-induced chondrocyte apoptosis and degradation of the extracellular matrix (ECM) play an important role in the progression of osteoarthritis (OA). In addition, tert-butylhydroquinone (TBHQ) is an activator of the nuclear factor erythroid derived-2-related factor 2 (Nrf2). The present study aimed to determine the effectiveness of TBHQ in preventing the apoptosis of chondrocytes and degradation of the extracellular matrix, induced by oxidative stress, in vitro. Therefore, rat chondrocytes were exposed to 20 μM tert-butyl hydroperoxide (TBHP) for 24 h to establish an oxidative damage model, in vitro. Thereafter, cell viability was evaluated using the Cell Counting Kit-8 assay. Moreover, the level of ROS was determined through 2′,7′-dichlorofluorescein diacetate staining. The mitochondrial membrane potential of chondrocytes was also measured using JC-1. Furthermore, cell apoptosis was assessed through Annexin V-fluorescein isothiocyanate/propidium iodide staining. The study also performed Western blotting and qPCR to evaluate the expression of extracellular matrix components, matrix catabolic enzymes, and changes in signalling pathways. The results showed that 2.5 and 5 μM of TBHQ reduced the TBHP-induced generation of excessive ROS and improved cell viability. Additionally, 2.5 and 5 μM of TBHQ prevented mitochondrial damage and apoptosis in rat chondrocytes. Treatment with TBHQ also increased the mRNA and protein expression levels of aggrecan and collagen II. However, TBHQ reduced the mRNA and protein expression levels of matrix metalloproteinase 3 (MMP3) and matrix metalloproteinase 13 (MMP13) in rat chondrocytes. In addition, treatment with TBHQ enhanced the protein expression levels of Nrf2, NADPH quinone oxidoreductase 1 (NQO-1), and hemeoxygenase-1 (HO-1) in rat chondrocytes. The current study showed that TBHQ was not only effective in protecting against TBHP-induced oxidative stress but also inhibited the apoptosis of rat chondrocytes and degradation of the ECM by activating the Nrf2 pathway. The results therefore suggest that TBHQ holds potential for use in the treatment of OA.

scholarly journals Estrogen receptor β agonist inhibits proliferation of endometrial stromal cells in an estrogen-receptor independent manner

2014 ◽  
Vol 66 (2) ◽  
pp. 735-741
Author(s):  
Qiong Xia ◽  
Dong Zhao ◽  
Juan Wang
Keyword(s):  
Estrogen Receptor ◽  
Protein Expression ◽  
Stromal Cells ◽  
Luciferase Reporter ◽  
Endometrial Stromal Cell ◽  
Independent Manner ◽  
Endometrial Stromal Cells ◽  
Mrna And Protein Expression ◽  
Ectopic Endometrium

The finding that estrogen receptor ER? agonists induce regression of ectopic endometrial implants in a rodent model has rekindled interest in the role of ER? in the growth and maintenance of human endometriosis. We hypothesize that the ER? agonist may have a direct effect on the human endometrium. We examined the mRNA and protein expression of ER? in both eutopic and ectopic endometrium and performed a luciferase reporter assay for the presence of functional ER? in human endometrial stromal cells. Diarylpropionitrile (DPN - a ER? specific agonist) significantly inhibited endometrial stromal cell proliferation at 10 M by 25% but not at 10 nM. ER? mRNA is present in both eutopic and ectopic endometrium; however, protein expression is absent in both stroma and epithelium. Our search for functional ER? similarly showed an absence in endometrial stromal cells. These results suggest that the regression of endometriosis might be ER?-independent.

scholarly journals 228.Progestin-induced proprotein convertase 6 is necessary for decidualisation of human endometrial stromal cells in vitro

2004 ◽  
Vol 16 (9) ◽  
pp. 228
Author(s):  
H. Okada ◽  
G. Nie ◽  
L. A. Salamonsen
Keyword(s):  
Real Time ◽  
Stromal Cells ◽  
Embryo Implantation ◽  
Functional Differentiation ◽  
Mrna Levels ◽  
Proprotein Convertase ◽  
Human Escs ◽  
Endometrial Stromal Cells ◽  
Mrna And Protein Expression

Decidualisation of human endometrium is an essential preparative event for successful establishment of pregnancy, and involves dramatic morphological and functional differentiation of the human endometrial stromal cells (ESCs). Proprotein convertase 6 (PC6) plays an important role in the processes of stromal cell decidualisation and embryo implantation in the mouse. PC6 is a member of the proprotein convertase family responsible for processing precursor proteins to their bioactive forms by selective proteolysis. In the present study we investigated the regulation of PC6 mRNA and protein expression in ESCs during decidualisation in vitro, and established a function for PC6 in decidualisation using morpholino antisense oligonucleotides (MOs). PC6 mRNA levels in ESCs during decidualisation were determined using quantitative real-time RT-PCR. 17β-oestradiol (E) plus medroxy-progesterone acetate (P) caused a significant increase in PC6 mRNA during decidualisation, whereas E alone did not increase PC6 mRNA expression. Consistent with the results of real-time PCR, much stronger PC6 immunostaining was observed in the cytoplasm of E plus P-treated ESCs (decidualised) compared to the E-treated ESCs (non-decidualised) on Day 12 of culture. This strong staining for PC6 was abolished by cotreatment with ZK 98299, a progesterone receptor antagonist. To investigate whether the induction of PC6 was necessary for decidualisation in vitro, MOs were used to block PC6 synthesis in cultured ESCs. PRL production, a typical marker for decidualisation, was significantly attenuated in decidualing ESCs following treatment with PC6 MOs in comparison to controls. These results suggest that PC6 plays a key role for decidualisation in human ESCs.

scholarly journals Upregulation of ETV2 Expression Promotes Endothelial Differentiation of Human Dental Pulp Stem Cells

2021 ◽  
Vol 30 ◽  
pp. 096368972097873
Author(s):  
Jing Li ◽  
Youming Zhu ◽  
Na Li ◽  
Tao Wu ◽  
Xianyu Zheng ◽  
...  
Keyword(s):  
Protein Expression ◽  
Dental Pulp ◽  
Tube Formation ◽  
Endothelial Differentiation ◽  
Cell Source ◽  
Lentiviral Infection ◽  
Mrna And Protein Expression ◽  
Human Dental Pulp

The lack of vasculogenesis often hampers the survivability and integration of newly engineered tissue grafts within the host. Autologous endothelial cells (ECs) are an ideal cell source for neovascularization, but they are limited by their scarcity, lack of proliferative capacity, and donor site morbidity upon isolation. The objective of this study was to determine whether differentiation of human dental pulp stem cells (DPSCs) into the endothelial lineage can be enhanced by recombinant ETV2 overexpression. DPSCs were extracted from fresh dental pulp tissues. ETV2 overexpression in DPSCs was achieved by lentiviral infection and cellular morphological changes were evaluated. The mRNA and protein expression levels of endothelial-specific markers were assessed through quantitative real-time polymerase chain reaction, western blot, immunofluorescence staining, and flow cytometry. The tube formation assay and Matrigel plug assay were also performed to evaluate the angiogenic potential of the ETV2-transduced cells in vitro and in vivo, respectively. Additionally, proteomic analysis was performed to analyze global changes in protein expression following ETV2 overexpression. After lentiviral infection, ETV2-overexpressing DPSCs showed endothelial-like morphology. Compared with control DPSCs, significantly higher mRNA and protein expression levels of endothelial-specific genes, including CD31, VE-Cadherin, VEGFR1, and VEGFR2, were detected in ETV2-overexpressing DPSCs. Moreover, ETV2 overexpression enhanced capillary-like tube formation on Matrigel in vitro, as well as neovascularization in vivo. In addition, comparative proteomic profiling showed that ETV2 overexpression upregulated the expression of vascular endothelial growth factor (VEGF) receptors, which was indicative of increased VEGF signaling. Taken together, our results indicate that ETV2 overexpression significantly enhanced the endothelial differentiation of DPSCs. Thus, this study shows that DPSCs can be a promising candidate cell source for tissue engineering applications.

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